Using Tetracysteine-Tagged TDP-43 with a Biarsenical Dye to Monitor Real-Time Trafficking in a Cell Model of Amyotrophic Lateral Sclerosis
Journal article, 2019

TAR DNA-binding protein 43 (TDP-43) has been identified as the major constituent of the proteinaceous inclusions that are characteristic of most forms of amyotrophic lateral sclerosis (ALS) and ubiquitin positive frontotemporal lobar degeneration (FTLD). Wild type TDP-43 inclusions are a pathological hallmark of >95% of patients with sporadic ALS and of the majority of familial ALS cases, and they are also found in a significant proportion of FTLD cases. ALS is the most common form of motor neuron disease, characterized by progressive weakness and muscular wasting, and typically leads to death within a few years of diagnosis. To determine how the translocation and misfolding of TDP-43 contribute to ALS pathogenicity, it is crucial to define the dynamic behavior of this protein within the cellular environment. It is therefore necessary to develop cell models that allow the location of the protein to be defined. We report the use of TDP-43 with a tetracysteine tag for visualization using fluorogenic biarsenical compounds and show that this model displays features of ALS observed in other cell models. We also demonstrate that this labeling procedure enables live-cell imaging of the translocation of the protein from the nucleus into the cytosol.

Author

Janice S.W. Ng

University of Cambridge

Maya A. Hanspal

University of Cambridge

Naunehal S. Matharu

University of Cambridge

Teresa P. Barros

University of Cambridge

Elin Esbjörner Winters

Chalmers, Biology and Biological Engineering, Chemical Biology

Mark R. Wilson

Illawarra Health and Medical Research Institute

University of Wollongong

Justin J. Yerbury

Illawarra Health and Medical Research Institute

University of Wollongong

C.M. Dobson

University of Cambridge

J.R. Kumita

University of Cambridge

Biochemistry

0006-2960 (ISSN) 1520-4995 (eISSN)

Vol. 58 39 4086-4095

Subject Categories

Cell and Molecular Biology

Biophysics

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1021/acs.biochem.9b00592

PubMed

31529970

More information

Latest update

11/11/2019